Peking University

Edigene Inc.

Chenxu Zhu

Chengqi Yi

Yuan Zhang

Wensheng Wei

Bo Xia

Shengjie Guo

Lulu Liu

RVDs with recognition preferences for 5mC, 5hmC and 6 mA and different binding properties to these epigenetic modifications are identified in this present invention. Methylation-dependent gene activation, efficient genome editing, targeted detection of 5hmC and other applications can be achieved by using these RVDs. The present invention therefore provides an isolated DNA binding polypeptide containing TALEs, a fusion protein, a polynucleotide, a vector comprising the polynucleotide and a host cell, and the use of the protein comprising TALE repeats domain in the preparation of a reagent for detecting a methylated base in a target sequence of a gene of interest, as well as a method for targeting and binding to a target sequence of a gene of interest in a cell.

Tale RVD specifically recognizing DNA base modified by methylation and application thereof

Disclosed are a method for marking 5-formyl cytosine and the use thereof in single base resolution sequencing. The method for marking the 5-formyl cytosine comprises the following steps of: (1) preparing a DNA or RNA sample; and (2) mixing the DNA or RNA sample with a buffer solution and a compound R&#x2014;CH&#x2014;CN to obtain a marking reaction system; and reacting the compound R&#x2014;CH&#x2014;CN therein with the 5-formyl cytosine in DNA and RNA molecules, and thereby achieving the marking of the 5-formyl cytosine; the reaction process is as in (I) below: wherein, Ris an electron withdrawing group next to the CHgroup, preferably &#x2014;CN, (II) or (III), and more preferably &#x2014;CN; R is a DNA or RNA molecule connected to the 5-formyl cytosine; and the pH value of the marking reaction system is 7.5-9. On this basis, also provided in the present invention is a sequencing analysis method for the 5-formyl cytosine. The method can be implemented at a single cell level, and can achieve the sequencing of single-base resolution levels.

Method for marking 5-formyl cytosine and use thereof in single base resolution sequencing

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Beijing, China

Chenxu Zhu